论文部分内容阅读
目的探讨复制缺陷型重组腺病毒载体(AdCMVeNOS)介导内皮型一氧化氮合酶(eNOS)基因转染对左向右分流所致的肺动脉高压及肺动脉高压危象兔的影响。方法 18只肺动脉高压模型兔分成实验组(n=9)和对照组(n=9),实验组气管内滴入AdCMVeNOS病毒转染液(5×109PFU/mL)2mL,对照组滴入生理盐水2mL。4d后结扎左向右分流,同时气管插管吸入10%氧气,检测两组肺动脉收缩压(SPAP)、肺动脉平均压(MPAP)和平均动脉压(MAP)的变化;同时采用硝酸还原酶法测定肺组织一氧化氮(NO)浓度,检测外源性eNOS基因mRNA的表达。结果实验组肺动脉压较转染前明显下降(P<0.01),而对照组肺动脉压无明显改变,与实验组比较,差异有统计学意义(P<0.05)。缺氧60min后,实验组仅4只兔出现肺动脉高压危象,而对照组全部出现肺动脉高压危象。两组缺氧后均出现氧分压(PaO2)降低,二氧化碳分压(PaCO2)升高,SPAP、MPAP升高,MAP降低。恢复供氧后,实验组各测得值恢复至正常水平,对照组SPAP和MPAP高于基础值,MAP低于基础值(P<0.01),实验组SPAP、MPAP和PaCO2的升高幅度以及MAP和PaO2的降低幅度均低于对照组(P<0.01)。免疫组化染色显示实验组肺泡内皮细胞、肺中小血管的平滑肌细胞和内皮细胞的eNOS表达均较对照组明显增加,而且肺组织的NO浓度也明显高于对照组(P<0.001)。琼脂糖凝胶电泳显示,实验组在3.77kb处扩增出特异条带,而对照组在3.77kb处未扩增出特异条带。结论 AdCMVeNOS转染左向右分流所致的兔肺动脉高压动物模型,可有效降低肺动脉压和肺动脉高压危象的发生率。
Objective To investigate the effect of replication-defective recombinant adenovirus vector (AdCMVeNOS) on endothelial-type inducible nitric oxide synthase (eNOS) gene transfection in rabbits with pulmonary hypertension and pulmonary hypertension induced by left-to-right shunt. Methods Eighteen rabbits with pulmonary hypertension were divided into experimental group (n = 9) and control group (n = 9). In the experimental group, 2 mL AdCMVNNV transfection solution (5 × 109 PFU / mL) 2mL. After 4 days, left-to-right shunt was ligated and 10% oxygen was inhaled through the endotracheal tube. The changes of pulmonary arterial pressure (SPAP), mean pulmonary artery pressure (MPAP) and mean arterial pressure (MAP) were measured. Nitric acid reductase Lung tissue nitric oxide (NO) concentration, detection of exogenous eNOS gene mRNA expression. Results Compared with the experimental group, the pulmonary arterial pressure in the experimental group decreased significantly (P <0.01), while the pulmonary arterial pressure in the control group did not change significantly. Compared with the experimental group, the difference was statistically significant (P <0.05). After 60 minutes of hypoxia, only 4 rabbits in the experimental group appeared pulmonary hypertension, while the control group all appeared the crisis of pulmonary hypertension. After the hypoxia, the oxygen partial pressure (PaO2) decreased, the partial pressure of carbon dioxide (PaCO2) increased, the levels of SPAP and MPAP increased and the MAP decreased. Recovery of oxygen, the experimental group measured values returned to normal levels, the control group SPAP and MPAP higher than the basic value, MAP was lower than the baseline value (P <0.01), experimental group SPAP, MPAP and PaCO2 increased amplitude and MAP And PaO2 decreased less than the control group (P <0.01). Immunohistochemical staining showed that the expression of eNOS in smooth muscle cells and endothelial cells of alveolar endothelial cells and small and large vessels in experimental group were significantly increased compared with control group, and the NO concentration in lung tissue was also significantly higher than that in control group (P <0.001). The agarose gel electrophoresis showed that the experimental group amplified a specific band at 3.77kb, while the control group did not amplify the specific band at 3.77kb. Conclusion AdCMVeNOS transfection of left pulmonary artery bypass grafting animal model of pulmonary hypertension can effectively reduce the incidence of pulmonary hypertension and pulmonary hypertension risk.